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<p>Tomato (<i>Solanum lycopersicum</i>) is an important horticultural crop and a widely used model for functional genomic research. However, stable genetic transformation in tomato remains a major experimental bottleneck. In this context, virus-based approaches have proven to be efficient alternatives for transient gene manipulation. In this study, we established and evaluated an integrated tobacco rattle virus (TRV)–based platform that combines virus-induced gene silencing (VIGS) and virus-mediated gene overexpression (VOX) in tomato. Using phytoene desaturase as a visual reporter, TRV-mediated VIGS demonstrated high efficiency, with photobleaching observed in over 95% of inoculated plants and sustained systemic silencing across vegetative and reproductive tissues. This phenotype was further supported by significant reductions in <i>SlPDS</i> transcript abundance and chlorophyll content. In parallel, the TRV-RNA2-pPEBV-MCS vector enabled robust VOX, as evidenced by strong systemic expression of green fluorescent protein at both the transcript and protein levels. Comparative analyses revealed higher VIGS and VOX efficiencies in <i>Nicotiana benthamiana</i> than in tomato, highlighting species-dependent differences in TRV performance. Overall, the optimized TRV-based VIGS and VOX systems provide complementary, efficient, and versatile tools for the rapid functional characterization of genes in tomato and related solanaceous species.</p>

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Tobacco Rattle Virus (TRV)-Mediated Gene Silencing and Overexpression in Tomato Plants via Agroinfiltration

  • 期刊论文

  • 发表日期：

    2026年2月10日

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  • Chae Woo Lim,
  • Junyoung Cho,
  • Dae Sung Kim,
  • Sung Chul Lee

摘要

Tomato (Solanum lycopersicum) is an important horticultural crop and a widely used model for functional genomic research. However, stable genetic transformation in tomato remains a major experimental bottleneck. In this context, virus-based approaches have proven to be efficient alternatives for transient gene manipulation. In this study, we established and evaluated an integrated tobacco rattle virus (TRV)–based platform that combines virus-induced gene silencing (VIGS) and virus-mediated gene overexpression (VOX) in tomato. Using phytoene desaturase as a visual reporter, TRV-mediated VIGS demonstrated high efficiency, with photobleaching observed in over 95% of inoculated plants and sustained systemic silencing across vegetative and reproductive tissues. This phenotype was further supported by significant reductions in SlPDS transcript abundance and chlorophyll content. In parallel, the TRV-RNA2-pPEBV-MCS vector enabled robust VOX, as evidenced by strong systemic expression of green fluorescent protein at both the transcript and protein levels. Comparative analyses revealed higher VIGS and VOX efficiencies in Nicotiana benthamiana than in tomato, highlighting species-dependent differences in TRV performance. Overall, the optimized TRV-based VIGS and VOX systems provide complementary, efficient, and versatile tools for the rapid functional characterization of genes in tomato and related solanaceous species.